Efficient, High-Quality\nEngineering of Therapeutic\nExtracellular Vesicles on an Integrated Nanoplatform

Engineered extracellular vesicles (EVs) have been recognized\nas\nimportant therapeutics for gene and cell therapy. To achieve clinically\ndesired therapy, technologies for EV engineering have high demands\non the efficacy in producing EVs and their qualities, which, however,\nremain challenging to conventional routes due to their limited control\non therapeutic payload delivery, EV secretion, and extracellular microenvironments.\nHere, we report a nanoplatform (denoted as PURE) that enables efficient\nelectro-transfection while stimulating cells to produce high-quality\nEVs carrying functional RNAs. PURE further employs an ammonium removal\nzone to maintain the physiological conditions of the extracellular\nmicroenvironment and an EV uptake zone that efficiently (87.1%) captures\nEVs in situ with porous hydrogels. The platform achieved about a 12-fold\nhigher yield of engineered EVs and a 146-fold abundance of desired\ntherapeutics compared to those naturally secreted from cells. The\nPURE-engineered miR-130a-EVs were validated for effectively upregulating\nthe mTOR signaling pathway in both in vitro and in vivo. Their therapeutic\ncapability was then verified by enhancing the in vitro activation\nof primordial follicles. In vivo applications further highlighted\nthe therapeutic effects of miR-130a-EVs in restoring ovary function\nin aged mice. The PURE platform represents a strategy for the clinical\ntranslation of EV-mediated therapy.